Disruption of the svkA gene encoding severin kinase.

Slides:

Advertisements

Similar presentations

Mutagenesis Methods Lily Peterson April 5 th, 2010.

Advertisements

Kristin Rosche, Emily Thornsen & Lloyd Turtinen  Department of Biology  University of Wisconsin-Eau Claire Knockout of the US29 gene of Human Cytomegalovirus.

How to get the cloned gene? (How to clone or identify the gene?) (1) From academic scientist (2) From company (3) PCR (RT-PCR) (4) From DNA library.

Spring 2016 BNFO 300 PTK7 protein domains involvement in PCP regulation of axon pathfinding of CoPA neurons in Zebrafish. Damien Islek:

Defining Epidermal Growth Factor Receptor exon 20 mutant sensitivity to tyrosine kinase inhibition Danny Rayes.

2470 bp 1891 bp WT bp 2314 bp A B Fig. S1. Verification with PCR amplification of the.

Supplemental Figure 2. (A) AtplaIVA-1 and AtplaIVA-2 null transcription lines for AtPLAIVA mRNA. RNAs from the relevant wild type Col were isolated.

Enhanced DNA damage in IDH1 mutant glioma cells.

Tamoxifen induced deletion of FAK

Vav‐1 gene‐targeting strategy.

The N‐terminus and SH2 domain of SOCS‐1 contribute to inhibition of JAK1 and JAK2 kinase activity. The N‐terminus and SH2 domain of SOCS‐1 contribute to.

Cloning of the t(1;5)(q23;q33) in a myeloproliferative disorder associated with eosinophilia: involvement of PDGFRB and response to imatinib by Kathryn.

Important scaffold function of the Janus kinase 2 uncovered by a novel mouse model harboring a Jak2 activation-loop mutation by Eric Keil, David Finkenstädt,

β-ARs signal cooperatively with mutant EGFR and inactivate LKB1

Volume 2, Issue 4, Pages (October 1998)

Expression and regulation of MD-2s.

AGL15 Interacts in a Sequence-Specific Manner with a Noncanonical CArG Motif Present in the Regulatory Region of AtGA2ox6.(A) Enrichment of the AtGA2ox6.

Molecular cloning of pms916 salt hypersensitive T-DNA mutant.

Targeted disruption of the mouse Pex14 gene.

BRCA2 Is Required for Homology-Directed Repair of Chromosomal Breaks

RAD51 is essential for L. donovani.

Adamtsl2 targeting strategy and validation of inactivation.

Analysis of GFP expression in gfp loss-of-function mutants.

APOE Gene Targeting (A) Schematic representation of the endogenous APOE locus, the gene targeting vector and the targeted APOE locus. The exons of the.

Impact of RTEL1 variants on telomere maintenance.

Cloning of a novel gene in the human kidney homologous to rat munc13s: Its potential role in diabetic nephropathy Yong Song, Menachem Ailenberg, Mel.

Cas9/gRNA-mediated gene deficiency in DF-1 cells.

Tagging of the endogenous Py03652 gene with the gfp gene in Plasmodium yoelii. Tagging of the endogenous Py03652 gene with the gfp gene in Plasmodium yoelii.

BRI1/BAK1, a Receptor Kinase Pair Mediating Brassinosteroid Signaling

Generation of Siva conditional knockout mice.

NILV‐S/MAR‐mediated transgene expression

Establishment of monoclonal SMN2-GFP reporter line in HEK293.

Expression of the mutants of HP1α and the mutant of SYCE2 in MCF7 cells or RPE cells. Expression of the mutants of HP1α and the mutant of SYCE2 in MCF7.

Dnmt2 mutants show delayed immune responses and Dnmt2–EGFP relocalizes and interacts with DCV RNA during infection. Dnmt2 mutants show delayed immune responses.

PCR amplification of the ORF encoding the cytosolic p36 protein of M

S. cerevisiae–E. coli chimeras have a partially rescued respiration-competent phenotype. S. cerevisiae–E. coli chimeras have a partially rescued respiration-competent.

Generation of Crb1-/- mice and confocal images of mouse retinas.

A highly conserved six-amino-acid region in the C-terminal CT domain of MARCH8 is responsible for its ability to downregulate TfR. A highly conserved six-amino-acid.

BAC recombineering, gene targeting and RMCE strategies.

Developmental defects in the svkA-null mutant.

PARP cleavage is detected in TA cells, but not in KSC

Geno- and phenotyping of selected keratinocyte cultures before and after conditional β-catenin gene ablation by Adenovirus-encoded Cre-recombinase. Geno-

TP53 western blot of primary cultured tail cells from both wild-type (WT) and Tp53Δ11/Δ11 (−/−) rats. TP53 western blot of primary cultured tail cells.

Fig. 1. Lack of Hmga1 and Hmga2 expression in A1/A2-KO mice

Expression of SvkA during development and induction of cytofission.

Characterisation of eSRS21 protein using an anti-eSRS21 antibody.

Fig. 1. Generation of WNK3 knockout mice

RXRa serves as a carrier for TR3 translocation initiated by 9-cis retinoic acid. RXRa serves as a carrier for TR3 translocation initiated by 9-cis retinoic.

HuD protein specifically recognizes and binds the Msi1 3′UTR sequence.

Fig. 4 Gene disruption via chip.

Analysis of Cdc7p at the end of meiosis.

The SIM domain of RAD51 is required for HR

Genomic structure of LTBP-4 around the 3rd 8-Cys repeat.

Gα12 is not required for hepatic cystogenosis induced by Pkd1 knockout

Amplicon sequencing analysis of on-target sites in trβ crispants

Kinetics of GFP secretion from CHO cells.

Targeted disruption of the mouse Atp1a2.

WT MARCH8, but not the CS mutant, specifically ubiquitinates and downregulates TfR. WT MARCH8, but not the CS mutant, specifically ubiquitinates and downregulates.

Vps36 interacts with Smo in the absence of Hh

Subnuclear localization of HA-tagged EBNA1 protein.

The dynamics of Akt activation in cultured human keratinocytes.

Increased electrophoretic mobility of Cx43 in the ischemic heart involves dephosphorylation at serine residues 325, 328 and/or 330. Increased electrophoretic.

Structure of the IFL1 Gene and the Nature of the Mutations in the ifl1 Alleles.(A) A schematic representation of the exon and intron organization of the.

Key functional sites of SPINDLIN1 could be phosphorylated by Aurora-A.

Competition between FLYWCH1/β-catenin and TCF4/β-catenin complexes for their interaction to Tcf-DNA-binding sites. Competition between FLYWCH1/β-catenin.

Construction of sgRNA expression cassette.

Genotype analysis of transient CRISPR system transformants.

RT-PCR Gel Blot Analyses of Cucurbita PP1 and PP2 mRNAs from Intergeneric Grafts of Cucumis sativus Scions on Cucurbita maxima or Cucurbita ficifolia Stocks.RT-PCR.

Cytokine expression induced by ligated-intestinal-loop assay using RT-PCR. Cytokine expression induced by ligated-intestinal-loop assay using RT-PCR. PCR.

Stable Cx43 expression in infected Cx43−/− cells and phosphorylation of Cx43. Stable Cx43 expression in infected Cx43−/− cells and phosphorylation of Cx43.

Presentation transcript:

Disruption of the svkA gene encoding severin kinase. Disruption of the svkA gene encoding severin kinase. (A) Two fragments of the kinase were amplified by PCR from genomic DNA, ligated to the blasticidin-resistance cassette and electroporated into AX2 wild-type cells. (B) Blasticidin-resistant clones were tested for knockout events by PCR1 (primers 1 and 2, 1385 bp for knockout) and PCR2 (primers 1 and 3, 2038 bp as a wild-type control). Under the conditions used, PCR2 did not ever amplify ∼3400 bp, including the blasticidin-resistance cassette, in the knockout, but the lack of a PCR product indicated a disruption of the svkA gene as well. (C) Western blot of 2×105 cells per lane, visualized with kinase-specific antibodies confirms the knockout at the level of SvkA protein and also shows the expression of the GFP fusion protein in the wild-type (first lane) and the mutant background (fourth lane). (D) GFP-SvkA-FL, immunoprecipitated from rescue cells, phosphorylated domains 2 and 3 of severin (DS211C, left), myelin basic protein (MyBP, right) and itself (both lanes). Meino Rohlfs et al. J Cell Sci 2007;120:4345-4354 © The Company of Biologists Limited 2007